大孔吸附树脂纯化美洲大蠊多肽的工艺研究

目的 筛选分离美洲大蠊多肽的最佳树脂,并优选其最佳纯化工艺。方法 以指标成分(多肽)定量检测结合体外肿瘤细胞抑制实验为评价指标,优选树脂类型并采用单因素试验考察影响纯化工艺的各种因素;采用氨基酸自动分析仪检测洗脱部位。结果 HP20分离效果最好,其最佳工艺参数为药液质量浓度0.3 g/m L,上样量2 BV,吸附体积流量2 BV/h,洗脱剂70%乙醇,洗脱剂用量2 BV,洗脱体积流量2 BV/h,多肽平均回收率为88.53%,洗脱部位含结合型氨基酸即多肽55.64%。结论 该工艺合理可行,分离效果好,可用于美洲大蠊抗肿瘤活性成分的富集与纯化。

Optimization in purification technique of polypeptides from Periplaneta american with macroporous adsorption resin

Objective To explore the optimal separation of polypeptides from Periplaneta americana by selecting appropriate macroporous resins and optimizing purification technology. Methods Taking the index component test and inhibition experiment of tumor cells in vitro method as indexes, macroporous resins type was chosen and single factor test was adopted to evaluate some factors affecting separating technology. The elution position was analyzed by Automatic amino acid analyzer. Results Macroporous resin HP20 had the best separating efficiency and the best purification conditions were as follows:the content in P. americana liquid 0.3 g/mL equivalent to raw material, the volume of drug 2 BV, the adsorption rate 2 BV/h, the volume of 70% ethanol as eluant 2 BV with elution rate of 2 BV/h. The recovery rate of polypeptides of elution position could reach 88.53% and the bound amino acids (polypeptides) was 55.64%. Conclusion This technology is reasonable and feasible and has good separation effect, which could be used for enrichment and purification of antineoplastic compounds from P. americana.